Endodormant grapevine buds require a period of chilling before they break and begin to grow. Custom Vitis bud cDNA microarrays (9,216 features) were used to examine gene expression patterns in overwintering Vitis riparia buds during 2,000 h of 4 degrees C chilling. Three-node cuttings collected concurrently with buds were monitored to determine dormancy status. Chilling requirement was fulfilled after 1,500 h of chilling; however, 2,000 h of chilling significantly increased the rate of bud break. Microarray analysis identified 1,469 significantly differentially expressed (p value < 0.05) array features when 1,000, 1,500, and 2,000 h of chilling were compared to 500 h of chilling. Functional classification revealed that the majority of genes were involved in metabolism, cell defense/stress response, and genetic information processing. The number of significantly differentially expressed genes increased with chilling hour accumulation. The expression of a group of 130 genes constantly decreased during the chilling period. Up-regulated genes were not detected until the later stages of chilling accumulation. Hierarchical clustering of non-redundant expressed sequence tags revealed inhibition of genes involved in carbohydrate and energy metabolism and activation of genes involved in signaling and cell growth. Clusters with expression patterns associated with increased chilling and bud break were identified, indicating several candidate genes that may serve as indicators of bud chilling requirement fulfillment.
BackgroundGenomic data release for the grapevine has increased exponentially in the last five years. The Vitis vinifera genome has been sequenced and Vitis EST, transcriptomic, proteomic, and metabolomic tools and data sets continue to be developed. The next critical challenge is to provide biological meaning to this tremendous amount of data by annotating genes and integrating them within their biological context. We have developed and validated a system of Grapevine Molecular Networks (VitisNet).Methodology/Principal FindingsThe sequences from the Vitis vinifera (cv. Pinot Noir PN40024) genome sequencing project and ESTs from the Vitis genus have been paired and the 39,424 resulting unique sequences have been manually annotated. Among these, 13,145 genes have been assigned to 219 networks. The pathway sets include 88 “Metabolic”, 15 “Genetic Information Processing”, 12 “Environmental Information Processing”, 3 “Cellular Processes”, 21 “Transport”, and 80 “Transcription Factors”. The quantitative data is loaded onto molecular networks, allowing the simultaneous visualization of changes in the transcriptome, proteome, and metabolome for a given experiment.Conclusions/SignificanceVitisNet uses manually annotated networks in SBML or XML format, enabling the integration of large datasets, streamlining biological functional processing, and improving the understanding of dynamic processes in systems biology experiments. VitisNet is grounded in the Vitis vinifera genome (currently at 8x coverage) and can be readily updated with subsequent updates of the genome or biochemical discoveries. The molecular network files can be dynamically searched by pathway name or individual genes, proteins, or metabolites through the MetNet Pathway database and web-portal at http://metnet3.vrac.iastate.edu/. All VitisNet files including the manual annotation of the grape genome encompassing pathway names, individual genes, their genome identifier, and chromosome location can be accessed and downloaded from the VitisNet tab at http://vitis-dormancy.sdstate.org.
Daylength is an important environmental cue for synchronizing growth, flowering, and dormancy with seasonality. As many floral development genes are photoperiod regulated, it has been suggested that they could have a regulatory role in bud endodormancy. Therefore, the influence of photoperiod was studied on inflorescence primordia differentiation and floral pathway related gene expression during the development of overwintering buds in Vitis riparia and V. spp. 'Seyval'. Photoperiod treatments were imposed 35 days after budbreak, and histological and transcriptomic analyses were conducted during the subsequent 42 days of bud development. Long day (LD, 15 h) and short day (SD, 13 h) buds were floral competent by 21 days of photoperiod treatment (56 days after budbreak); however, the floral meristem developed faster in LD than in SD buds. Analysis of 132 floral pathway related genes represented on the Affymetrix Grape Genome array indicated 60 were significantly differentially expressed between photoperiod treatments. Genes predominantly related to floral transition or floral meristem development were identified by their association with distinct grape floral meristem development and an expression pattern in LD consistent with their previously identified roles in flowering literature. Genes with a potential dual role in floral development and dormancy transitioning were identified using photoperiod induced differences in floral development between LD and SD buds and uncharacteristic gene expression trends in relation to floral development. Candidate genes with the potential to play a dual role in SD dormancy induction include circadian rhythm or flowering transition related genes: AP2, BT1, COL-13, EIN3, ELF4, DDTR, GAI and HY5.
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